Detangling the Web: Isolation of the MaSp1 Promoter from the Black Widow spider
Poster Number
13
Format
Poster Presentation
Faculty Mentor Name
Craig Vierra
Abstract/Artist Statement
The long-term objective of this research experiment is to understand how the expression of silk genes is regulated in black widow spiders. Syuthetic silk production will be possible in the near future but understanding how a spider expresses these genes is one of the most critical steps. In this research proposal, we attempted to clone the promoter region of the fibroin MaSp 1 from the black widow spider. Different regions of the MaSpl promoter were amplified using polymerasechain reaction using different primers that were designed to amplify the 5' -flanking QNA region of the MaSpl gene. We were able to successfully amplify promoter regions sized at 300 bp, 1000 bp-and 1500 bp. After amplification of the different promoter regions from genomic DNA, we inserted the fragments into the cloning vector pBAD-TOPO. Acquisition of the promoter fragments were confirmed by restriction digestion and agarose gel electrophoresis. Our future goal is to insert these fragments into a promoterless reporter plasmid and measure the transcriptional activity of these DNA regions in insect cells.
Location
Wendell Phillips Center, 1st floor hallways
Start Date
3-5-2008 1:00 PM
End Date
3-5-2008 3:00 PM
Detangling the Web: Isolation of the MaSp1 Promoter from the Black Widow spider
Wendell Phillips Center, 1st floor hallways
The long-term objective of this research experiment is to understand how the expression of silk genes is regulated in black widow spiders. Syuthetic silk production will be possible in the near future but understanding how a spider expresses these genes is one of the most critical steps. In this research proposal, we attempted to clone the promoter region of the fibroin MaSp 1 from the black widow spider. Different regions of the MaSpl promoter were amplified using polymerasechain reaction using different primers that were designed to amplify the 5' -flanking QNA region of the MaSpl gene. We were able to successfully amplify promoter regions sized at 300 bp, 1000 bp-and 1500 bp. After amplification of the different promoter regions from genomic DNA, we inserted the fragments into the cloning vector pBAD-TOPO. Acquisition of the promoter fragments were confirmed by restriction digestion and agarose gel electrophoresis. Our future goal is to insert these fragments into a promoterless reporter plasmid and measure the transcriptional activity of these DNA regions in insect cells.